CRISPR/Cas-based precision gene replacement in plants via microhomology-mediated end joining DOI Open Access
Tien Van Vu, Gah‐Hyun Lim,

Seung Hee Choi

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Aug. 27, 2022

Abstract Precise gene or allele replacement is a desirable technology, but implementing it in plants remains challenging. CRISPR-Cas-based approaches, such as targeting (GT) and prime editing (PE), have opened up new possibilities for precise plants. However, their size efficiency still need improvement. Recently, strategies using canonical nonhomologous end-joining (cNHEJ) microhomology-mediated end joining (MMEJ) been considered promising alternatives yeast mammals. these approaches not extensively explored applied to Here, we proposed tested tool, termed PREMJ ( p recision re placement via m icrohomology-mediated joining), precision The strategy employing 20-bp microhomology MMEJ donors (∼100 bp lengths) inhibitor, NU7441, produced high targeted efficiencies, 1.60 ± 0.14, 4.47 1.98, 8.98 4.73 % protoplasts of tomato, lettuce, cabbage, respectively. Our data also revealed the critical impacts length NU7441 concentration on PREMJ-based Although obtaining edited challenging due inefficient protoplast regenerations Agrobacterium -mediated delivery, may significantly contribute competent complex delivery plant regeneration. Key Message We designed method, PREMJ, CRISPR-Cas-mediated double-stranded break (DSB) formation repairing DSBs with donor template carrying desired base changes microhomologies flanking DSB ends. showed feasibility tomatoes, albeit its efficacy transformation requires further optimization.

Language: Английский

Precise deletion, replacement and inversion of large DNA fragments in plants using dual prime editing DOI Creative Commons

Yidi Zhao,

Zhengwei Huang,

Ximeng Zhou

et al.

Nature Plants, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 13, 2025

Language: Английский

Citations

3
Engineering source-sink relations by prime editing confers heat-stress resilience in tomato and rice DOI

H. F. Lou,

Shujia Li,

Zihang Shi

et al.

Cell, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 1, 2024

Language: Английский

Citations

11
Optimized dicot prime editing enables heritable desired edits in tomato and Arabidopsis DOI
Tien Van Vu,

Ngan Thi Nguyen,

J.-G. Kim

et al.

Nature Plants, Journal Year: 2024, Volume and Issue: 10(10), P. 1502 - 1513

Published: Sept. 6, 2024

Language: Английский

Citations

10
Recent Advances in Tomato Gene Editing DOI Open Access
Eduardo Larriba, О. M. Yaroshko, José Manuel Pérez‐Pérez

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(5), P. 2606 - 2606

Published: Feb. 23, 2024

The use of gene-editing tools, such as zinc finger nucleases, TALEN, and CRISPR/Cas, allows for the modification physiological, morphological, other characteristics in a wide range crops to mitigate negative effects stress caused by anthropogenic climate change or biotic stresses. Importantly, these tools have potential improve crop resilience increase yields response challenging environmental conditions. This review provides an overview techniques used plants, focusing on cultivated tomatoes. Several dozen genes that been successfully edited with CRISPR/Cas system were selected inclusion illustrate possibilities this technology improving fruit yield quality, tolerance pathogens, responses drought soil salinity, among factors. Examples are also given how domestication wild species can be accelerated using generate new better adapted climatic situation suited indoor agriculture.

Language: Английский

Citations

6
Adapting crops for climate change: regaining lost abiotic stress tolerance in crops DOI Creative Commons
Michael Palmgren,

Sergey Shabala

Frontiers in Science, Journal Year: 2024, Volume and Issue: 2

Published: Dec. 5, 2024

It is often stated that agricultural outputs need to increase substantially meet the demands for more food posed by a growing population. However, when accounting climate change, we argue current projected increases are unrealistic and realistic goal would be maintain yields per area of production. This will require breeding crops with increased tolerance abiotic stresses such as drought, salinity, waterlogging, high temperatures. can accomplished in one two ways: introducing stress genes into present high-yielding or increasing already tolerant orphan and/or wild plants. We first strategy easing restrictions on use gene editing technologies making substantial improvements cell-based phenotyping identify available pool crop its relatives. The success second depend number domestication selected order obtain comparable present-day cultivars. still too early conclude which strategies, rewilding (bringing lost from ancestors back domesticated crops) de novo (domesticating resilient plants underutilized directly), most effective future sustainable agriculture. given importance issue, some rapid action needs taken.

Language: Английский

Citations

5
How to use CRISPR/Cas9 in plants: from target site selection to DNA repair DOI Creative Commons
Adéla Přibylová, Lukáš Fischer

Journal of Experimental Botany, Journal Year: 2024, Volume and Issue: 75(17), P. 5325 - 5343

Published: April 22, 2024

Abstract A tool for precise, target-specific, efficient, and affordable genome editing is a dream many researchers, from those who conduct basic research to use it applied research. Since 2012, we have that almost fulfils such requirements; based on clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems. However, even CRISPR/Cas has limitations obstacles might surprise its users. In this review, focus the most frequently used variant, CRISPR/Cas9 Streptococcus pyogenes, highlight key factors affecting mutagenesis outcomes: (i) activity, as effect of target sequence, chromatin state, or Cas9 how long remains in place after cleavage; (ii) follow-up DNA repair mechanisms including mostly cell type cycle phase, but also, example, ends produced by cleavage (blunt/staggered). Moreover, note some differences between using plants, yeasts, animals, knowledge individual kingdoms not fully transferable. Awareness these can increase likelihood achieving expected results plant editing, which provide detailed guidelines.

Language: Английский

Citations

4
GENOME EDITING FOR IMPROVING DISEASE RESISTANCE IN RICE (Oryza sativa L.) - SCOPE, ACHIEVEMENTS, LIMITATIONS AND FUTURE NEEDS DOI
Shobica Priya Ramasamy,

Sakthi Ambothi Rathnasamy,

Sudha Manickam

et al.

Physiological and Molecular Plant Pathology, Journal Year: 2025, Volume and Issue: unknown, P. 102640 - 102640

Published: March 1, 2025

Language: Английский

Citations

0
Molecular tools for genome editing in plants: a synthetic overview DOI Creative Commons
Nicolau Brito da Cunha, Michel Lopes Leite, Júlio Carlyle Macedo Rodrigues

et al.

Vegetable Research, Journal Year: 2025, Volume and Issue: 5(1), P. 0 - 0

Published: Jan. 1, 2025

Language: Английский

Citations

0
Integration of CRISPR/Cas9 with multi-omics technologies to engineer secondary metabolite productions in medicinal plant: Challenges and Prospects DOI

Anupriya Borah,

Sumit Kumar Singh,

Rituja Chattopadhyay

et al.

Functional & Integrative Genomics, Journal Year: 2024, Volume and Issue: 24(6)

Published: Nov. 4, 2024

Language: Английский

Citations

2
Progress and prospect of genome editing tools development and molecular breeding in cotton DOI Open Access

Guangqin Yang,

Guanying Wang,

Lu Yu

et al.

Chinese Science Bulletin (Chinese Version), Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 1, 2024

Genome editing technology can achieve efficient and accurate at the biological gene transcription level, which is a revolutionary in field of life science. The genome system based on Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein (CRISPR/Cas), has become most widely used plants with its simple, fast characteristics. Cotton an important economic crop main cultivated species upland cotton (G. hirsutum L.) allotetraploid plant complex genome. contains At- Dt- two subgenomes multiple copy, therefore, functional redundancy common. Therefore, conventional T-DNA insertion chemistry, physical mutagenesis other technologies to create mutants are very low efficiency cotton. In recent years, Chinese researchers have established tools including CRISPR/Cas9/12a/12b, CBE/ABE/ABE8e/CABE base editor, Cas13a/b/c dCas9-TV transcriptional activation, knock out, in, down, up point mutation target genes. addition, they evaluate off-target effects whole re-sequencing strategy. Whether be effectively delivered step editing, genotype receptor material plays decisive role effective delivery genetic transformation mainly mediated by Agrobacterium. Jin668, high efficiency, world present, for exploring function endogenous genes conducting research. Here, Successive Regeneration Acclimation (SRA) strategy obtain Jin668 analysis regeneration mechanism introduced detail. It expected break genotypic restriction varieties during regeneration. this review, establishment process particular. Using above tools, created mutant library more than 5,000 genes, fiber quality, resistance disease, insect, temperature, salt alkali, seed haploid induction traits, promoting development basic research molecular breeding This paper also discusses limitations itself problems solutions. At end paper, future direction prospected. era 4.0, namely stage design breeding, trend future. For cotton, "precise genome+efficient editing" drive

Language: Английский

Citations

1